新型 CCCH 型锌指蛋白 ZC3H4 在二氧化硅诱导的上皮间质转化中的新兴作用。

The emerging roles of a novel CCCH-type zinc finger protein, ZC3H4, in silica-induced epithelial to mesenchymal transition.

机构信息

Department of Physiology, School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, China; Department of Respiration, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, China; Department of Pharmacology, School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, China; Key Laboratory of Environmental Medicine Engineering, Ministry of Education, School of Public Health, Southeast University, Nanjing, Jiangsu, 210009, China.

Department of Physiology, School of Medicine, Southeast University, Nanjing, Jiangsu, 210009, China.

出版信息

Toxicol Lett. 2019 Jun 1;307:26-40. doi: 10.1016/j.toxlet.2019.02.014. Epub 2019 Mar 1.

DOI:10.1016/j.toxlet.2019.02.014

PMID:30826420

Abstract

BACKGROUND

The epithelial to mesenchymal transition (EMT) contributes to fibrosis during silicosis. Zinc finger CCCH-type containing 4 protein (ZC3H4) is a novel CCCH-type zinc finger protein that activates inflammation in pulmonary macrophages during silicosis. However, whether ZC3H4 is involved in EMT during silicosis remains unclear. In this study, we investigated the circular ZC3H4 (circZC3H4) RNA/microRNA-212 (miR-212) axis as the upstream molecular mechanism regulating ZC3H4 expression and the downstream mechanism by which ZC3H4 regulates EMT as well as its accompanying migratory characteristics.

METHODS

The protein levels were assessed via Western blotting and immunofluorescence staining. Scratch assays were used to analyze the increased mobility induced by silica. The CRISPR/Cas9 system and small interfering RNAs (siRNAs) were employed to analyze the regulatory mechanisms of ZC3H4 in EMT and migration changes.

RESULTS

Specific knockdown of ZC3H4 blocked EMT and migration induced by silicon dioxide (SiO). Endoplasmic reticulum (ER) stress mediated the effects of ZC3H4 on EMT. circZC3H4 RNA served as an miR-212 sponge to regulate ZC3H4 expression, which played a pivotal role in EMT. Tissue samples from mice and patients confirmed the upregulation of ZC3H4 in alveolar epithelial cells.

CONCLUSIONS

ZC3H4 may act as a novel regulator in the progression of SiO-induced EMT, which provides a reference for further pulmonary fibrosis research.

摘要

背景

上皮间质转化（EMT）有助于矽肺纤维化。锌指 CCCH 型结构域 4 蛋白（ZC3H4）是一种新型的 CCCH 型锌指蛋白，在矽肺中激活肺巨噬细胞炎症。然而，ZC3H4 是否参与矽肺中的 EMT 尚不清楚。在这项研究中，我们研究了环状 ZC3H4（circZC3H4）RNA/微小 RNA-212（miR-212）轴作为调节 ZC3H4 表达的上游分子机制，以及 ZC3H4 调节 EMT 及其伴随的迁移特征的下游机制。

方法

通过 Western blot 和免疫荧光染色评估蛋白质水平。划痕实验用于分析二氧化硅诱导的迁移增加。CRISPR/Cas9 系统和小干扰 RNA（siRNA）用于分析 ZC3H4 在 EMT 和迁移变化中的调节机制。

结果

ZC3H4 的特异性敲低阻断了二氧化硅（SiO）诱导的 EMT 和迁移。内质网（ER）应激介导了 ZC3H4 对 EMT 的影响。circZC3H4 RNA 作为 miR-212 的海绵，调节 ZC3H4 的表达，在 EMT 中发挥关键作用。来自小鼠和患者的组织样本证实了 ZC3H4 在肺泡上皮细胞中的上调。